Human eccrine sweat contains tissue kallikrein and kininase II

We attempted to determine the level of sweat kallikrein (kininogenase) and to purify and characterize it using sweat collected over a white petrolatum barrier. Thermally induced eccrine sweat obtained from 24 healthy subjects showed kallikrein activity of 24.4 ng kinins generated/1 mg of sweat protein when heated plasma was used as the substrate and 16.1 ng kinin when purified low molecular weight bovine kininogen was used as the substrate. Sweat was sequentially purified by Sephacryl S-200, diethyaminoethyl Sephacel, and fast flow liquid chromatography Mono Q chromatography. Sweat kallikrein had a M(r) of 40,000 and was inhibited by aprotinin but not by soybean trypsin inhibitor. The peptide generated by sweat kallikrein was identified as lys-bradykinin using reverse phase high-performance liquid chromatography and by its amino acid sequence. Anti-human urinary kallikrein immunoglobulin G neutralized the sweat kallikrein activity completely, indicating that the sweat kallikrein is the glandular type. Purified sweat and salivary kallikrein showed similar M(r) and responses to inhibitors and antibodies. Using immunohistochemistry, kallikrein activity was localized in luminal ductal cells and in the peripheral rim of secretory coil segments, presumably the outer membrane of the myoepithelium. We also observed kininase activity in sweat at M(r) 160,000, which was inhibited by ethylenediamine tetraacetic acid, captopril, and angiotensin converting enzyme inhibitor peptide, indicating that it is kininase II (or angiotensin converting enzyme). Sweat also contains abundant non-kallikrein hydrolases for S-2266 and S-2302. The demonstration of glandular kallikrein, its tissue localization, and the presence of kininase II in sweat provide the basis for future studies on the physiologic role of the kallikrein/kinin system in the eccrine sweat gland.